The long term objectives of proposed study is to produce transgenic/knockout(KO) rabbits as model animals for human cardiovascular disease (CVD) and other diseases. Rabbit has been long recognized a better CVD model animal than mouse and rat, however transgenic/KO rabbit production is hampered largely by low efficiency rate of the conventional DNA microinjection method and the unavailability of embryonic stem cell lines. We propose a novel solution to aforementioned problems: use genetically manipulated somatic cells for nuclear transfer (NT) to produce transgenic/KO clones. In Phase I study, we choose C-reactive protein (CRP) as the targeted gene. We propose to produce CRP knockout cloned rabbits. The specific aims include: (1) identify 2-3 well-characterized rabbit fibroblast cell lines; (2) genetically manipulate these cell lines at different passages, generating CRP knockout cell lines; (3) produce cloned rabbit embryos using KO cell lines vs. non-KO cell lines; and (4) transfer cloned embryos to produce CRP-KO-NT rabbits. These efforts will establish the foundation for commercially producing transgenic/KO rabbits as model animals for CVD and other human disease studies. Such transgenic rabbit facility is not available in the United States at this moment and we believe our proposed project will for the first time make rabbit, the valuable however under-studied model animal, available to researchers throughout the country and worldwide. [unreadable] [unreadable] [unreadable]